Gene Therapy

Patent metadata

Field	Value
Publication number	US-2025288617-A1
Application number	US-202318859829-A
Country	US
Kind code	A1
Filing date	Apr 28, 2023
Priority date	Apr 29, 2022
Publication date	Sep 18, 2025
Grant date	—

Abstract

Use of a combination of (a) cyclosporin H (CsH) or a derivative thereof, and (b) a p53 inhibitor and/or an adenoviral protein, or one or more nucleotide sequences encoding therefor, for increasing the efficiency of gene editing of an isolated population of cells when transduced by a viral vector and/or increasing the efficiency of transduction of an isolated population of cells by a viral vector.

First claim

1 . Use of a combination of (a) cyclosporin H (CsH) or a derivative thereof, and (b) a p53 inhibitor and/or an adenoviral protein, or one or more nucleotide sequences encoding therefor, for increasing the efficiency of gene editing of an isolated population of cells when transduced by a viral vector and/or increasing the efficiency of transduction of an isolated population of cells by a viral vector.

2 . A method of transducing a population of cells comprising the steps of:
i) contacting the population of cells with a combination of (a) cyclosporin H (CsH) or a derivative thereof, and (b) a p53 inhibitor and/or an adenoviral protein, or one or more nucleotide sequences encoding therefor; and ii) transducing the population of cells with a viral vector.

3 . The method according to claim 2 , further comprising the step of iii) introducing a site-directed nuclease into the cells.

4 . The method according to claim 3 , wherein the site-directed nuclease is a CRISPR/Cas9 nuclease.

5 . The use or method according to any preceding claim , wherein the combination comprises cyclosporin H (CsH) or a derivative thereof, a p53 inhibitor and an adenoviral protein.

6 . The use or method according to any preceding claim , wherein the p53 inhibitor is a p53 dominant negative peptide.

7 . The use or method according to any preceding claim , wherein the p53 inhibitor is GSE56 or a variant thereof; pifithrin-a or a derivative thereof; or an siRNA, shRNA, miRNA or antisense DNA/RNA.

8 . The use or method according to any preceding claim , wherein the p53 inhibitor is GSE56 or a variant thereof.

9 . The use or method according to any preceding claim , wherein the p53 inhibitor comprises or consists of an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 3.

10 . The use or method according to any preceding claim , wherein the adenoviral protein is from an Adenovirus of serotype 5.

11 . The use or method according to any preceding claim , wherein the adenoviral protein is E4ORF6/7.

12 . The use or method according to any preceding claim , wherein the adenoviral protein comprises or consists of an amino acid sequence having at least 70% to SEQ ID NO: 27.

13 . The use or method according to any preceding claim , wherein the one or more nucleotide sequences are in the form of mRNA.

14 . The use or method according to any preceding claim , wherein the viral vector is an Integration-Defective Lentiviral Vector (IDLV).

15 . The use or method according to any preceding claim , wherein the cells are haematopoietic stem and/or progenitor cells (HSPCs).

16 . The use or method according to any preceding claim , wherein the cells are T cells.

17 . The use or method according to any preceding claim , wherein the cells are contacted with cyclosporin H prior to and/or at the same time as contact with the viral vector.

18 . The use or method according to any preceding claim , wherein the cells are contacted with a p53 inhibitor and/or an adenoviral protein, or one or more nucleotide sequences encoding therefor, prior to and/or at the same time as contact with a site-directed nuclease.

19 . The method according to any one of claims 3 to 18 , wherein contact with the site-directed nuclease occurs about 10-14 hours, optionally about 12 hours after the beginning of the transduction step.

20 . The use or method according to any preceding claim , wherein the population of cells is transduced by the viral vector in a single transduction step or in two transduction steps.

21 . A combination of (a) cyclosporin H (CsH) or a derivative thereof, and (b) a p53 inhibitor and/or an adenoviral protein, or one or more nucleotide sequences encoding therefor, for use in a method of gene editing.

22 . A population of transduced cells, prepared according to the method of any one of claims 2 to 20 .

23 . A pharmaceutical composition comprising the population of transduced cells of claim 22 .

24 . A method of treating or preventing a disease in a subject, comprising administering the population of cells according to claim 22 , or composition according to claim 23 , to the subject.

25 . A population of cells according to claim 22 , or a composition according to claim 23 , for use in a method of treating or preventing a disease in a subject.

26 . A method of gene therapy comprising the steps:
(a) transducing a population of cells according to the method of any one of claims 2 to 20 ; and (b) administering the transduced cells to a subject.

Assignees

• Ospedale San Raffaele Srl
• Fond Telethon Ets

Inventors

• Ferrari Samuele
• Jacob Aurelien
• Naldini Luigi

Classifications

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N2501/26

  Flt-3 ligand (CD135L, flk-2 ligand) · CPC title

• C12N2501/22

  Colony stimulating factors (G-CSF, GM-CSF) · CPC title

• C12N2501/145

  Thrombopoietin [TPO] · CPC title

• C12N2501/125

  Stem cell factor [SCF], c-kit ligand [KL] · CPC title